Illumina MiSeq测序平台测定蒙古羊瘤胃液相和固相菌群多样性  

Bacteria Community Diversity of Liquid and Solid Phases of Ruminal Contents of Mongolian Sheep Analyzed by Illumina Mi Seq Platform

作　　者：曾燕[1,2] 简平[1,2] 倪学勤[1,2] 周毅[1,2] 祝辉[1,2] 曾东[1,2] 

机构地区：[1]四川农业大学动物医学院动物微生态研究中心,雅安625014 [2]动物疫病与人类健康四川省重点实验室,雅安625014

出　　处：《动物营养学报》2015年第10期3256-3262,共7页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：教育部留学回国人员科研启动基金资助项目(200835-1);四川省学术带头人培养基金资助项目

摘　　要：本研究旨在比较蒙古羊瘤胃液相和固相细菌菌群多样性。应用Illumina Miseq测序平台的16S r RNA基因扩增子技术对采集自5只蒙古羊的瘤胃液相和固相样品进行高通量测序,采用UPARSE-操作分类单元(OTU)法和非加权组平均(UPGMA)法分析试验结果。结果表明,共获得91 971条序列和15 321个OTU,平均每个样品有13 138条序列和2 189个OTU。液相与固相样品相比:门水平分类上存在显著性差异的优势菌群为拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)(P<0.05),拟杆菌门在液相和固相样品中的比例分别为67%和45%;属水平分类上存在显著性差异的优势菌群为普氏菌属(Prevotella)和1种未鉴定(unknown)的属(P<0.05),液相样品优势菌群为普氏菌属,所占比例为53%,而固相样品优势菌群来自1种未鉴定的属,所占比例为32%;种水平分类上存在显著性差异的是栖瘤胃普雷沃菌(Prevotella ruminicola)(P<0.05),在液相和固相样品中所占的比例分别为41%和11%。与液相样品相比,固相样品的alpha多样性较好;主成分分析(PCA)和主坐标分析(PCo A)显示来自同一动物个体的瘤胃液相和固相样品聚类在一起。结果提示,蒙古羊瘤胃液相和固相样品均具有丰富的细菌菌群;与液相样品相比,固相样品细菌的多样性更好。This experiment was conducted to compare bacteria community diversity of liquid and solid phases of ruminal contents of Mongolian sheep. The samples of liquid and solid phases of ruminal contents from 5 Mongolian sheep were investigated using 16 S r RNA amplicon high-throughput sequencing technique by Illumina Mi Seq platform,and the results were analyzed by UPARSE-operational taxonomic units（ OTU） and unweighted pair group method with arithmetic mean（ UPGMA）. The results showed that a total of 91 971 sequences and 1 871 OTUs across all samples were observed,and the average number of sequence and OTU was 13 138 and 267 per sample,respectively. Compared between the samples of liquid and solid phases： at phyla level,dominant bacteria with significant differences were Bacteroidetes and Firmicutes,and the percentage of Bacteroidetes of the samples of liquid and solid phases was 67% and 45%,respectively; at genus level,dominant bacteria with significant differences were Prevotella and an unknown genus,dominant bacterium of liquid phase was Prevotella whose percentage reached 53%,and that of solid phase was the unknown genus whose percentage reached 32%;at species level,dominant bacteria with significant differences were Prevotella ruminicola,and the percentage of liquid and solid phases reached 41% and 11%,respectively. The alpha diversity of solid phase was higher compared with that of liquid phase,and the samples from the same animal clustered together by principal component analysis（ PCA） and principal coordinate analysis（ PCo A）. In conclusion,both the contents of liquid and solid phases have high diversity of bacteria,and the diversity of solid phase is higher compared with that of liquid phase.

关 键 词：ILLUMINA MiSeq 液相 固相 细菌 多样性 

分 类 号：S826[农业科学—畜牧学]